...Origin of Replication? See our Plasmids 101: Origin of Replication blog).  The replication problem Plasmids...directly inhibit replication by binding to the origin of replication and blocking replication machinery (del...cell have the same replicon and/or partitioning system. Let’s start with the replicon- the part of the ...plasmid that contains the origin of replication and the replication control machinery (Need a refresher... with the same replicon are incompatible because they compete for the same replication control machinery... that inhibits replication when copy number in the cell is high, but allows replication when copy number...either indirectly inhibit replication through inhibiting translation of replication machinery proteins, or...

...ori2, ori3) and replication initiation proteins (RepA, RepB) necessary for replication in various hosts...the ori and allow for replication. However, if plasmids have their own replication machinery such as Rep... and W are known to have replicons with broad host ranges. Popular replicons from these groups include...different hosts that a plasmid can transfer to, replicate in, and persist in. Some plasmids have a narrow... while others have a broad host range and can replicate and stably maintain the genes they carry among.... These features are: The plasmid origin of replication (ori) contains structural elements that are versatile...structure allows plasmids to utilize diverse host replication machinery. For instance, the ori typically contains...

...because devaluing science has alarming implications for our public policies—many of which impact reproductive...science to the public, talking to policy-makers, and broadly defending science in the public sphere as often...Canada.  Outside the lab, she dabbles in science policy, public outreach, and restorative justice.  Follow ...restrictions that conflict with scientific evidence. We need representatives and policymakers who not only ...approach policy makers take in abusing climate science skepticism to promote harmful policies that benefit...move slowly, news about possible broad changes to policies seems to break everyday. Many researchers and ...participation in government, evidence to inform policies, a protected place to work, funding for science...

.... Useful for nuclear isolation and scRNA-seq applications. Jonathan Ting Viviana Gradinaru 163505-PHPeB...

...the grant application process. When my PhD advisor first asked me to write a grant application due the ...the next round of applications. Step 5: Keep writing! Each written grant application builds upon the ... project grant application" by the NIH Still want more information on the application process? What types...advisor alone applies for these competitive grant applications; however, in some cases, your advisor may choose...writing, volunteered to take the lead on various applications throughout my graduate career, and helped receive... Step 1: Your PI forwards you a Request For Applications (RFA), what next? First, check the submission...that if a PI asks for assistance in the grant application process, she has faith in you! Additionally, ...

...are not compatible with regular mesophilic systems are the psychrophilic enzymes.  The reason behind this...weather. Psychrophilic enzymes face very similar issues when forced to express in mesophilic host systems...about placing the mesophilic hosts at 0°C? That won’t work out either - most mesophilic systems would either...mediated editing for psychrophilic systems as well as engineering and regulating psychrophilic recombinant proteins... on a special class of enzymes expressed by psychrophilic (cold-loving) bacteria. These bacteria inhabit...inhabit cold temperature locales and psychrophilic enzymes help them survive under severely cold conditions...with a few basics.  Mesophiles vs. extremophiles Mesophilic organisms are your regular next-door microbes...

...in PubMed and choose the appropriate publication. This publication will be listed at the top of the final...6) or you can click the "Save" button and jump around to the different pages by clicking on the appropriate...Add Plasmids page and click the blue "Submit Deposit" button. Figure 13: Click the "Submit Deposit" button... account, you can begin the deposit process by clicking "Submit Online Deposit" (for online submission... menu: Figure 1: Begin the deposit process by clicking "Submit Online Deposit" or "Download Deposit Spreadsheet...submitting plasmid(s) using a spreadsheet (right), click the “Download Deposit Spreadsheet” button to download... "Published plasmids" or "Unpublished or pre-publication plasmids" and set a desired distribution status...

...and attending seminars in bioethics, public health, science policy, and science and technology studies ...number of organizations in public engagement efforts, including giving public lectures, writing science...developments in genetics research, their applications and regulatory policies; keeping tabs on the conversation...path to science policy and science communication A career in science communication or policy requires broad...scientists arguably have a duty to engage the public on the implications of their work. Johnny Kung is the Director... space of science education, communication, and policy. In this post, I hope to tell you a little more... as well as the personal, social and ethical implications of genetics. In these conversations, pgEd strives...

...physiological performance.  2. Trans-splicing Overview: For trans-plicing, splice site donor and acceptor sequences...followed by a splice site donor. The AAV encoding part B of the transgene contains a splice site acceptor...can enhance the rate of trans-splicing (Lai et al., 2005). Trans-splicing does not always outperform single...either the overlapping or the trans-splicing mechanism. The splice site donor and acceptor sequences and...sequence, upstream of the 5’ splice donor and downstream of the 3’ splice acceptor of the transgene to...part B of the transgene form a heterodimer, then splicing of the pre-mRNA of the two halves of the transgene...the full-length transgene.  Drawbacks:  Trans-splicing occurs across the ITR junction, and this is likely...

...and autism, and other hypotheses mount yet policies and public opinion do not seem to take note. For example...to inform the public (8–13). Facing the possibility that reason is losing ground in public discourse, many...Democrats more often than Republicans, the March is not about Democrats or Republicans (16). This is a Pro-...inclusion of science to inform policy (17). The facts surrounding a matter or policy should have more weight ... Idea Pew Research Center: Public Praises Science; Scientists Fault Public, Media March for Science: Principles...when we drop them. Scientific realities limit what policy can do and, in this way, science has always been...what outcomes are actually possible for a given policy. In exchange, the government protects (and often...

...Technical Tips). Minigenome replication and transcription In order to replicate the minigenome, the viral...transcription and replication strategies of Marburg virus and Ebola virus by using artificial replication systems...allowing researchers to study aspects of the EBOV replication cycle under BSL-2 conditions (4). What is a ...that are crucial cis-acting signals for genome replication and viral gene transcription (6). The EBOV genome...5’ ends of the genome which are required for replication and transcription (Figure 1). All viral genes...cis-acting signals necessary to direct minigenome replication and reporter gene transcription by the EBOV polymerase... in 2, 5). The minigenome is capable of being replicated and transcribed in the cell when the appropriate...

...good quality application. Even following this advice, only about 50% of my grant applications have been ...do need to be able to research and write your application, then give it to someone else to read and / or...it). It’s easy for reviewers to spot when an application is rushed (whether the idea isn’t properly researched...researched / developed or the application is full of typos) and these are unlikely to get past the initial...someone else in the department) to read over your application and give you feedback. They’ve most likely been...alterations before submitting - handing them your application 3 days before the deadline isn’t going to be ...problem in a novel way. Finally, engineering new applications to solve problems will provide originality to...

...science is all about navigating conflict. In literary terms, conflict can be defined as (1) the struggle...story, you need to have conflict. There are a lot of ways to introduce conflict into your work, but one...problem?” Those are the conflicts that will drive your story forward.  Beyond the Conflict  Once you have your...that the audience can understand the conflict. A science-based conflict gives you an easy framework with ...presence of obstacles that need to be overcome. Conflict, and its inherent tension, is interesting. It’...invested in seeing how they do it.    Finding Conflict in Science  In all science communication, what...talk to a scientist about their research, these conflicts come out quite naturally as they explain their...

...Background noise Probe Allelic specificity Can be multiplexed Single amplicon validation technique ... and dsDNA. This is not true for all amplicons; if your amplicon is AT-rich or has a secondary structure...understanding and technical proficiency with common applications - now let’s dive right in! You know qPCR is...as it solves, so understanding and using the application correctly is imperative. Picking your chemistry...hairpin design of probes/primers often offer higher allelic specificity, due to the need for the target:probe...Bustin, 2017 for a more in-depth review.)  Your amplicon should be between 70-200 bp long With ΔΔCT analysis... intensity should smoothly increase as the DNA amplicon transitions from single-stranded to double-stranded...

...offer export license applications for VSV and SARS-COV-1 plasmids. Export control license process    An...excited to announce we now offer an export license application for rabies materials! Some of the materials...cannot expedite the application process.  More information about export control licenses is available through...the USA, where Addgene is located.  The export license, if granted, allows us to ship rabies plasmids ...  An export control license, once approved, allows Addgene to ship plasmids or viral vectors containing...will need to provide a brief description of the applicable research project when purchasing export-controlled...export-controlled materials. Addgene will then apply for the license for distribution to the requester’s organization...

...  Limitations of the in silico approach The in silico approach cannot replicate the complexity of antigen... risk of an antibody.  In silico prediction of adverse reactions In silico assessment uses computer modeling...from the in silico assessments, as well as analyze the contributing factors that in silico prediction ... drugs are still immunogenic, meaning that they elicit an immune response from the body. Several factors...responses vary depending on the person. However, in silico and in vitro assessment of immunogenicity can help...drug epitopes likely to be presented by T cells to elicit an immune response. T cell epitopes are found in... evaluate the immunogenicity of therapeutics in silico. These algorithms are trained on vast amounts of...

...received 116,224 employment based green card applications and approved 91,023 during the first 3 quarters... development, business development, consulting, policy, big data and the list goes on and on. As you think...options discussed in this post benefit from the applicant thinking about the process early, keeping good...occupation" as requiring theoretical and practical application of a body of highly specialized knowledge in ...bachelor's degree or its equivalent and state licensure, if required to practice in that field. H-1B work-authorization...Hire American" Executive Order which sets broad policy intentions directing federal agencies to propose... April 1 deadline for companies to submit an application on their behalf.  If an individual obtains a ...

...that in a bit! Viral genomes and replication When it comes to replication, viruses tend to march to the ...methods of replication. In fact, an entire classification scheme, known as the Baltimore replication classes...developed around how they replicate (Figure 2). Figure 2: Baltimore replication classes. Image credit: ...an intermediary DNA replication step between DNA and mRNA. DNA genome replication can involve an RNA or...cell. The cell then uses its own machinery to replicate the viral genome and proteins. This is, for the...before being translated into proteins. RNA viruses replicate their genomes using a variety of RNA or DNA intermediaries... bacteria and eukaryotes, which use a binary replication process whereby one cell copies itself and “splits...

...Lentiviral, miRNA expression (PRIME) system for application in knockdown of gene expression at a single copy...

...after credible attempts to replicate major findings could not be replicated by other researchers.There...published research is difficult to replicate.  Why is replicating previous research findings so difficult...can be difficult to replicate. Unclear methods Until someone attempts to replicate the results of another...positives in original studies and false negatives in replications are unavoidable, but can be minimized with high...obtaining a different result after conducting a replication study. An important difference between the original... methods, QRPs, and small samples, then when replications do not come out the same as an original research...data, materials, and code Only when you try to replicate the work of other researchers (or your own from...